Sepsis is an often fatal clinical syndrome that develops after infection or injury. Sepsis is the most frequent cause of mortality in hospitalized patients. Experimental models of gram negative sepsis based on administration of bacterial endotoxin (lipopolysaccharide, LPS) have led to an improved understanding of the pathogenic mechanisms of lethal sepsis and conditions related to sepsis by virtue of the activation of a common underlying inflammatory cytokine cascade. This cascade of host-response mediators includes TNF, IL-1, PAF and other macrophage-derived factors that have been widely studied as acute, early mediators of eventual lethality in severe endotoxemia (Zhang and Tracey, In The Cytokine Handbook, 3rd ed. Ed Thompson (Academic Press Limited, USA). 515-547, 1998).
Unfortunately, therapeutic approaches based on inhibiting these individual “early” mediators of endotoxemia have met with only limited success in large prospective clinical trials against sepsis in human patients. It is possible to infer from these disappointing results that later-appearing factors in the host response might critically determine pathogenesis and/or lethality in sepsis and related disorders. Accordingly, there is a need to discover such putative “late” mediators necessary and/or sufficient for part or all of the extensive multisystem pathogenesis, or for the lethality, of severe endotoxemia, particularly as endotoxemia is representative of clinical sepsis and related clinical disorders.
HMG1 is a 30 kDa chromosomal nucleoprotein belonging to the burgeoning high mobility group (HMG) of non-histone chromatin-associated proteins. As a group, the HMG proteins recognize unique DNA structures and have been implicated in diverse cellular functions, including determination of nucleosome structure and stability, as well as in transcription and/or replication. The HMG proteins were first characterized by Johns and Goodwin as chromatin components with a high electrophoretic mobility in polyacrylamide gels (see in The HMG Chromosomal Proteins, E. W. Johns, Academic Press, London, 1982). Higher eukaryotes exhibit three families of HMG proteins: the HMG-1/-2 family, the HMG-14/-17 family and the HMG-1/-Y family. Although the families are distinguishable by size and DNA-binding properties, they are similar in their physical properties. HMG proteins are highly conserved across species, ubiquitously distributed and highly abundant, and are extractable from chromatin in 0.35 M NaCl and are soluble in 5% perchloric or trichloroacetic acid. Generally, HMG proteins are thought to bend DNA and facilitate binding of various transcription factors to their cognate sequences, including for instance, progesterone receptor, estrogen receptor, HOX proteins, and Oct1, Oct2 and Oct6. Recently, it has become apparent that a large, highly diverse group of proteins including several transcription factors and other DNA-interacting proteins, contain one or more regions similar to HMG1, and this feature has come to be known as the HMG1 box or HMG1 domain. cDNAs coding for HMG1 have been cloned from human, rat, trout, hamster, pig and calf cells, and HMG1 is believed to be abundant in all vertebrate cell nuclei. The protein is highly conserved with interspecies sequence identities in the 80% range. In chromatin, HMG1 binds to linker DNA between nuclesomes and to a variety of non-β-DNA structures such as palindromes, cruciforms and stem-loop structures, as well as cisplatin-modified DNA. DNA binding by HMG1 is generally believed to be sequence insensitive. HMG1 is most frequently prepared from washed nuclei or chromatin, but the protein has also been detected in the cytoplasm. (Reviewed in Landsman and Bustin, BioEssays 15:539-546, 1993; Baxevanis and Landsman, Nucleic Acids Research 23:514-523, 1995). To date, no link has been established between the HMG proteins and any clinical condition or disease.
HMG1 has been alternatively identified as a heparin-binding protein abundantly expressed in developing brain and dubbed “amphoterin” for its highly dipolar sequence, comprising two internal repeats of a positively charged domain of about 80 amino acids (the HMG1 box) and an acidic C-terminal domain containing a stretch of approximately 30 continuous glutamic or aspartic acid residues. Amphoterin/HMG1 has been localized to the outer surface of the plasma membranes of epithelial, and especially neuronal cells, where it has been specifically localized to the filipodia of neural cells. Inhibition studies have suggested that amphoterin/HMG1 is required for process (neurite) extension and amphoterin/HMG1 also may be involved in neuron-glia interactions (Merenmies et al., J. Biol. Chem. 266:16722-16729, 1991; Merenmies et al., J. Biol. Chem. 266:16722-16729, 1991; Milev et al., J. Biol. Chem. 273:6998-7005, 1998; and Salmivirta et al., Exp. Cell Res. 200:444-451, 1992). Amphoterin/HMG1 can be released from murine erthroleukemia cells after stimulation with the chemical inducer hexamethylenebisacetamide (Melloni et al., Biochem. Biophys. Res. Commun. 210:82-89, 1995). Previous study suggested that the gene product of the HMG1 gene functions as a differentiation enhancing factor by stimulating α-PKC (Melloni et al., Biochem. Biophys. Res. Commun. 210:82-89, 1995; and Melloni et al., FEBS Lett. 368:466-470, 1995).
The HMG1 gene product has been shown to interact with plasminogen and tissue-type plasminogen activator (t-PA) and effectively enhance plasmin generation at the cell surface, a system that is known to play a role in extracellular proteolysis during cell invasion and tissue remodeling. Amphoterin/HMG1 has also been shown to interact with the receptor of advanced glycosylation end products (RAGE) (Mohan et al., Biochem. Biophys. Res. Commun. 182:689-696, 1992; Yamawaki et al., J. Neurosci. Res. 44:586-593, 1996; Salmivirta et al., Exp. Cell Res. 200:444-451, 1992; and Vassalli et al., J. Clin. Invest. 88:1067-1072, 1991), (Redlitz and Plow, Baillieres Clin. Haematol. 8:313-327, 1995; and Parkkinen et al., J. Biol. Chem. 266:16730-16735, 1991).
There is a longstanding need in the art to discover improved agents that can prevent the cytokine-mediated inflammatory cascade and have therapeutic activity in a large variety of cytokine-mediated inflammatory diseases. The present invention was made during the course of investigative research to identify agents that mediate toxicity, pathogenesis and/or lethality in sepsis and other disorders related by a common activation of the inflammatory cytokine cascade.
Disease and conditions mediated by the inflammatory cytokine cascade are numerous.
Such conditions include the following grouped in disease categories:
                Systemic Inflammatory Response Syndrome, which includes:                    Sepsis syndrome            Gram positive sepsis            Gram negative sepsis            Culture negative sepsis            Fungal sepsis            Neutropenic fever            Urosepsis                        Menin gococcemia        Trauma hemorrhage        Hums        Ionizing radiation exposure        Acute pancreatitis        Adult respiratory distress syndrome (ARDS)Reperfusion Injury, which includes        Post-pump syndrome        Ischemia-reperfusion injuryCardiovascular Disease, which includes        Cardiac stun syndrome        Myocardial infarction        Congestive heart failureInfectious Disease, which includes        HIV infection/HIV neropathy        Meningitis        Hepatitis        Septic arthritis        Peritonitis        Pneumonia Epiglottitis        E. coli 0157:H7            Hemolytic uremic syndrome/thrombolytic thrombocytopenic purpura    Malaria    Dengue hemorrhagic fever    Leishmaniasis    Leprosy    Toxic shock syndrome    Streptococcal myositis    Gas gangrene    Mycobacterium tuberculosis    Mycobaclerium aviun intracellulare     Pneumocystis carinii pneumonia    Pelvic inflammatory disease    Orchitis/epidydimitis    Legionella    Lyme disease    Influenza A    Epstein-Barr Virus    Viral associated hemiaphagocytic syndrome    Viral encephalitis/aseptic meningitisObstetrics/Gynecology, including:            Premature labor        Miscarriage        InfertilityInflammatory Disease/Autoimmunity, which includes:        Rheumatoid arthritis/seronegative arthropathies        Osteoarthritis        Inflammatory bowel disease        Systemic lupus erythematosis        Iridoeyelitis/uveitistoptic neuritis        Idiopathic pulmonary fibrosis        Systemic vasculitis/Wegener's gramilomatosis        Sarcoidosis        Orchitis/vasectomy reversal proceduresAllergic/Atopic Diseases, which includes:        Asthma        Allergic rhinitis        Eczema        Allergic contact dermatitis        Allergic conjunctivitis        Hypersensitivity pneumonitisMalignancy, which includes:        ALL        AML        CML        CLL        Hodgkin's disease, non-Hodgkin's lymphoma        Kaposi's sarcoma        Colorectal carcinoma        Nasopharyngeal carcinoma        Malignant histiocytosis        Paraneoplastic syndrome/hypercalcemia of malignancyTransplants, including:        Organ transplant rejection        Graft-verus-host disease            CachexiaCongenital, which includes:            Cystic fibrosis        Familial hematophagocytic lymphohistiocytosis        Sickle cell anemiaDermatologic, which includes:        Psoriasis        AlopeciaNeurologic, which includes:        Multiple sclerosis        Migraine headacheRenal, which includes:        Nephrotic syndrome        Hemodialysis        UremiaToxicity, which includes:        OKT3 therapy        Anti-CD3 therapy        Cytokine therapy        Chemotherapy        Radiation therapy        Chronic salicylate intoxicationMetabolic/Idiopathic, which includes:        Wilson's disease        Hemachromatosis        Alpha-1 antitrypsin deficiency        Diabetes        Hashimoto's thyroditis        Osteoporosis        Hypothalamic-pituitary-adrenal axis evaluation        Primary biliary cirrhosis        